Lactate Dehydrogenase

Lactate Dehydrogenase
Lactate Dehydrogenase (LDH) is an important enzyme in humans. It occurs in different regions of the body, each region having a unique conformation of different subunits. The Jmole image shown is that of LDH-5, the form found in skeleton muscle and the liver. LDH is a key enzyme in anaerobic respiration. Anaerobic Respiration is the conversion of pyruvate into lactate acid in the absence oxygen. This pathway is important to glycolysis in two main ways. The first is that if pyruvate were to build up glycoysis and thus the generation of ATP would slow. The second is anaerobic respiration allows for the regeneration of NAD+ from NADH. NAD+ is required when glyceraldehyde-3-phosphate dehydrogenase oxidizes glyceraldehyde-3-phosphate in glycolysis, which generates NADH. Lactate dehydrogenase is responsible for the anaerobic conversion of NADH to NAD+.

Structure
LDH is a quaternary protein formed of the combination of two subunits, M and H (Muscle and Heart) into a structure of four of the subunits. The various combinations found in the human body are:
 * (4H) Heart
 * (3H1M) Reticuloendothelial
 * (2H2M) Lungs
 * (1H3M) Kidneys
 * (4M) Muscle and Liver

The secondary structure of LDH as shown here is comprised of 40% alpha helices and 23% beta sheets. The SCOP data classifies this form of lactate dehydrogenase as mixed beta-alpha-beta, with mainly parallel beta sheets.

Catalysis
Studies have shown that the reaction mechanism of LDH follows an ordered sequence. In order for lactate to be oxidized NADH must bind to the enzyme first followed by lactate. Several residues are involved in the binding of NADH, including Lys 250 and Tyr 85. Once the NADH is bound to the enzyme, it is then possible for lactate to bind. Lactate binds to the enzyme between the nicotinamide ring and His 195. Transfer of a hydride ion then happens quickly in either direction giving a mixture of the two tertiary complexes, enzyme-NAD+-lactate and enzyme-NADH-pyruvate .Finally pyruvate dissociates from the enzyme followed by NAD+.



Kinetics
Kinetic studies of lactate dehydrogenase with oxalate and oxamate (structural analogues of lactate and pyruvate)have proven the mechanism stated above. The rate limiting step in this reaction is the rate of dissociation of NAD+ and NADH. The conversion of pyruvate to lactate with the subsequent regeneration of NAD+ is very favorable.



Regulation
As the mechanism is one of equilibrium, There appears to be no regulation specifically designed for lactate dehydrogenase, instead it is dependent on the activation of anaerobic reparation and the presence of pyruvate and NADH, or lactate and NAD+.

3D structures of lactate dehydrogenase
2e37, 2v6m – TtL-LDH – Thermus thermophilus 2xxe – TtL-LDH (mutant)  2x8l, 1ceq – PfL-LDH – Plasmodium falciparum 2zqy, 2zqz, 1llc – L-LDH – Lactobacillus casei 3d0o - SaL-LDH – Staphylococcus aureus 2v65 - L-LDH A chain – Champsocephalus gunnari 2v6b – L-LDH – Deinococcus radiodurans 2frm – CpLDH – Cryptosporidium parvum 1sov, 1pze - TgL-LDH – Toxoplasma gondii 1v6a – L-LDH A chain – Cyprinus carpio 1y6j – L-LDH – Clostridium thermocellum 1ldb - GsL-LDH – Geobacillus stearothermophilus<BR /> 1ldm, 6ldh, 8ldh – sdM4-LDH – spiny dogfish<BR /> 2ldx – C4-LDH - mouse

LDH binary complex
2ydn – PfL-LDH + bicine<BR /> 1cet - PfL-LDH + chloroquine<BR /> 1t2c - PfL-LDH + NAD<BR /> 1ez4 - L-LDH + NAD – Lactobacillus pentosus<BR /> 1lld, 2ldb - GsL-LDH + NAD<BR /> 2a92 – PvL-LDH + NAD – Plasmodium vivax <BR /> 2a94, 2aa3 - PvL-LDH + NAD-analog

1u4o, 1u4s, 1u5a, 1xiv – PfL-LDH + inhibitor<BR /> 2xxb – TtL-LDH (mutant) + AMP

L-LDH ternary complex with inhibitor
1u5c - PfL-LDH + inhibitor + NAD<BR /> 1t24, 1t25, 1t26 - PfL-LDH + azole derivative + NAD<BR /> 1t2e - PfL-LDH (mutant) + oxamate + NAD<BR /> 1ldg - PfL-LDH + oxamate + NAD<BR /> 2xxj – TtL-LDH (mutant) + oxamate + NAD<BR /> 2v7p - TtL-LDH + oxamate + NAD<BR /> 3om9, 3czm – TgL-LDH + OXQ + NAD<BR /> 3h3f – L-LDH A chain + oxamate + NAD – rabbit<BR /> 2fnz - CpLDH + oxamate + NAD<BR /> 1t2f - hL-LDH B chain + azole derivative + NAD – human<BR /> 1i0z - hL-LDH H chain + oxamate + NAD<BR /> 1i10 - hL-LDH M chain + oxamate + NAD<BR /> 1oc4 – L-LDH + oxamate + NAD – Plasmodium berghei<BR /> 1a5z - L-LDH + oxamate + NAD – Thermotoga maritima<BR /> 1lth - L-LDH + oxamate + NAD – Bifidobacterium longum<BR /> 1ldn - GsL-LDH + oxamate + NAD<BR /> 9ldb, 9ldt - pL-LDH + oxamate + NAD – pig

L-LDH ternary complex with reactants
1sow, 1pzh - TgL-LDH + oxalate + NAD<BR /> 1t2d - PfL-LDH + oxalate + NAD<BR /> 1pzf - TgL-LDH + oxalate + NAD-analog

1pzg - TgL-LDH + sulfate + NAD-analog

3h3j – SaL-LDH (mutant) + pyruvate + NAD<BR /> 3d4p - SaL-LDH + pyruvate + NAD<BR /> 2fn7 - CpLDH + lactate + NAD<BR /> 2ewd - CpLDH + pyruvate + NAD-analog

2fm3 - CpLDH + pyruvate + NAD<BR /> 5ldh - pLDH + citrate + NAD-analog

3ldh - sdLDH + pyruvate + NAD<BR />

D-LDH
3kb6 – D-LDH + lactate + NAD – Aquifex aeolicus<BR /> 1j49 – LdD-LDH + sulfate + NAD – Lactobacillus delbrueckii<BR /> 1j4a - LdD-LDH + sulfate<BR /> 1f0x – D-LDH + FAD – Escherichia coli<BR /> 2dld - D-LDH + oxamate + NAD – Lactobacillus helveticus

L-LDH II
1x0a, 1vbi – TtL-LDH II

Additional Information
For additional information, see Carbohydrate Metabolism

Reference

 * 1- http://www.bioc.aecom.yu.edu/labs/calllab/highlights/LDH.htm
 * 2- http://www.cheric.org/ippage/e/ipdata/2004/05/file/e200405-701.pdf
 * 3- http://resources.metapress.com/pdf-preview.axd?code=ulnhp23038060m21&size=largest
 * 4- http://www.u676.org/Documents/Chretien-ClinChimActa-95.pdf
 * 5- http://www.jbc.org/content/243/17/4526.full.pdf+html